Regulation of Invertebrate Biological Control Agents in Europe: Recommendations for a Harmonised Approach

Abstract

There have been few reported negative effects associated with the import and release of non-native invertebrate biological control agents (IBCAs), yet this practice is subject to stringent regulation in a number of countries including the USA, Canada, Australia and New Zealand. The import and release of IBCAs in Europe is not regulated by an EU Directive, as is the case for microorganisms and semiochemicals. As a consequence, some European countries have regulatory systems, others do not, and among countries with regulation, there is no consistency in the information requirements that biocontrol companies must produce when seeking a licence to release non-native species. Against this background, the REBECA Action: 1. Produced a standardised Application Form for the licenced release of non-native IBCAs in Europe, together with an accompanying Guidance Document. 2. Recommended adoption of a step-wise testing procedure for the environmental risk assessment of insect, mite and nematode agents, and summarised methods to assess establishment, host range and dispersal. 3. Endorsed the reactivation and updating of the EPPO ‘Positive List’. 4. Proposed the establishment of an Expert Group to provide advice on the first release in Europe of non-native IBCAs. This chapter reviews the scientific and political dimensions underlying these recommendations and proposals, and provides an update toward the objective of a harmonised regulatory framework for non-native IBCAs in Europe.

1 Appendix 1: Application Form for the Import, Shipment, Rearing and Release of Invertebrate Biological Control Agents in European Countries Guidance on the completion of this Application form is provided in a separate document – Appendix 2.

Guidance on the completion of this Application form is provided in a separate document – Appendix 2.

Guidance on the completion of this Application form is provided in a separate document – Appendix 2.

Using this Form

This form should be used for the submission of an application to a National Competent Authority (NCA) of the European Union (EU) for a permit to license the import for research, mass-rearing and/or release of an invertebrate natural enemy used for the biological control of invertebrate and plant pests (Invertebrate Biological Control Agent or IBCA) and for other beneficial organisms. Organisms include invertebrates as well as entomopathogenic nematodes,Footnote

See REBECA WP 5 – Recommendations for regulation requirements for entomopathogenic nematodes

but not micro-organisms. Guidance on the completion of this form is provided in the accompanying Guidance Document¹. This form is valid for an application relating to a single biological control organism. An organism is characterised as any identifiable and recognisable taxon of the IBCA, either a species, or recognised sub-species, population, strain or biotype.

After the NCA has received your application (administrative forms and documentation/dossier), you will receive an acknowlegdement of receipt within a specified period of time. The application will then be checked for completeness and subjected to a risk assessment in relation to the purpose of your application (e.g. for research under quarantine conditions, or a commercial release). The risk analysis will be conducted by the NCA or – upon its request – by a specified expert or group of experts. The NCA will conduct a risk analysis in the light of the information provided, or any other sources they have available. The NCA may need to contact you to clarify parts of the application or to seek further information. At all times and in all communication, including that with external experts, your application will be regarded as confidential. After the risk assessment has been completed, the NCA will make a decision as to whether to grant a permit within a previously agreed period of time. The licence to permit an import and/or release will be valid for a fixed period of time, assigned by the NCA, after which a renewal may be sought, or a request may be made to place the organism on the EPPO Positive List. In the case of mixed products, an application should be made for each separate component.

Information Required to Complete this Form

This application form and related information requirements for the release of non-indigenous IBCAs contains 5 parts (numbered 1–5) and is structured in a step-wise way: depending on the origin of the organism and the purpose of the application, the sequence of assessments and level of information required is related to the perceived level of risk. An application for any specified organism should include the following information:

Part 1. Application information

• A. Information on the applicant

• B. Purpose of the application and use

Part 2. Information for indigenous and non-indigenous IBCAs

• A. Taxonomy and origin

• B. Product information

Where an application is made for the import for research and rearing of a non-indigenous species and/or release of a native IBCA, the applicant should proceed to sections 4 and 5 of the form. Where the application is for the release of a non-indigenous IBCA, section 3 of this form must be completed.

Part 3. Information requirements for intentional release of a non-indigenous IBCA with reference to:

• A. Biology and ecology

• B. Assessment of risks and benefits

• a. Establishment,

• b. Host specificity

• c. Dispersal

• d. Direct and indirect effects

Part 4. Submission of forms and Signature

• A. Submission details

• B. Agreement: safeguards and signature

Part 5. Appendices

Sections of the Form to be Completed

This form can be used for the import and release of all IBCAs. Depending on the purposes of use, either some or all parts of the form must to be completed.

• 1. Renewal of a previous application Parts 1, 4 and 5

• 2. First application

• Organism on Positive List Parts 1, 2, 4 and 5

• Import only Parts 1, 2, 4 and 5

• Release of indigenous IBCAs Parts 1, 2, 4 and 5

• Release of non-indigenous IBCAs Parts 1, 2, 3, 4 and 5

For more information: Call... or refer to our website...... or consult the Guidance Document

2 Part I. Application Information

Table 1

Table 2

Table 3

Table 4

3 Part 2. Information for Indigenous and Non-indigenous IBCAs

Table 5

Table 6

Table 7

In the case of a renewal of a previously successful application (Section 1.3), or if the species or population is indigenous to the country or ecoregion, and/or imported for research or rearing only and/or is mentioned on the list of species considered safe for use in the intended area of release, no further information is required and only the submission details in 4A and B and Appendices (Part 5) need to be completed. For other applications, such as the release of a non-indigenous species, the information requirements in Part 3 must be supplied.

4 Part 3. Information Requirements for Intentional Release of a Non-indigenous IBCA

Table 8

Table 9

Table 10

5 Part 4. Submission of Forms and Signature

Table 11

Table 12

4.3 General Safeguards

The applicant or authorized user undertaking the release proceeds under the conditions of the authorization for release, taking into account of the following requirements:

• All appropriate safety procedures should be put in place.

• Any relevant information on adverse effects, which might relate to the released IBCA, should be reported immediately to the National Competent Authority (NCA).

• Information on sites and dates of supply or release of the IBCA should be made available to the NCA, if requested.

• Information requirements have been supplied according to the most recent knowledge, and that the conditions made by the NCA will be respected.

Table 13

6 Part 5. Appendices

1 Appendix 2: Guidelines for the Completion of an Application for the Import, Shipment, Rearing and Release of Invertebrate Biological Control Agents in European Countries These guidelines are largely based on Bigler et al., 2005 and redrafted during REBECA workshop discussions in 2005–2007.

These guidelines are largely based on Bigler et al., 2005 and redrafted during REBECA workshop discussions in 2005–2007.

These guidelines are largely based on Bigler et al., 2005 and redrafted during REBECA workshop discussions in 2005–2007.

Using this Guidance

The purpose of this document is to provide guidance on how to complete the application form for a permit for the import (including labelling, packaging and storage in transit), mass-rearing and/or release of an Invertebrate Biological Control Agent (IBCA) and other beneficial organisms.Footnote

IPPC, 2005 – http://www.ippc.int/: Any organism directly or indirectly advantageous to plants or plant products,

including biological control agents [ISPM No. 3, 2005, ISPM No. 5, 2007])

The application form and this accompanying guidance document are intended to cover all situations in which a permit (licence) is required: (1) for import and release, (2) for species and strains, (3) for different types of biological control programmes (augmentative, classical biocontrol, weeds) and includes, (4) product and efficacy information. The environmental risk assessment (ERA) and risk/benefit analysis will be based upon the information provided in the application form. It is therefore important that all required parts of the form are completed. It is also recommended that all EU countries should use the same application and guidance documents. The National Competent Authority (NCA) will conduct a risk analysis in the light of the information provided, or any other sources they have available. The dossier to be submitted to the NCA must include information on the organismFootnote

Organism = any identifiable taxon of an IBCA; either a species, recognised sub-species, population, strain or biotype. Natural enemy = predator, parasitoid or EPN known to attack and develop on a certain host or prey and intended to be used for the biological control of certain plants, plant pests, stored products; IBCA = product of a certain specified natural enemy; non-indigenous = organism (taxon) orginated and collected ouitside the area of release. For other terminology, the IPPC definitions are used.

(IBCA) for import (including shipment), research, rearing and/or release as specified in the following parts of the application form:

• Part 1. Information on the applicant (A) and purpose of the application and use (B)

• Part 2. Information on the invertebrate biological control agent: identity, specific characteristics, origin and distribution (A), and product information (B)

• Part 2. Information relating to intentional release of a non-indigenous IBCA: biology and ecology of the IBCA (A) and an assessment of risks and benefits of the release (B)

• Part 4. Information on where to send the application (A) and conditions (B)

• Part 5. Appendices

Parts 1–5 of this guidance document are divided into different sections and sub-sections. The title and number of each part, section and sub-section referred to in this document correspond with the same parts, sections and sub-sections of the application form. In the case of renewal of an application, Parts 1, 4 and 5 have to be completed. In the case of a first application, Parts 1, 2, 4 and 5 must be completed by all applicants, including applications for the release of indigenous species, when required by the NCA. For applications to release a non-indigenous species, Part 3 of the application form must also be completed.

For more information: Call... Or check our website......

2 Information to Be Submitted by the Applicant

3 Part 1. Application Information

Table 14

Provide information (including contact details) on:

• 1.1. Who will apply for the permit⁴; include confirmation of the person’s authorization and a copy of a valid identification card with the application.

• 1.2. The contact person, research manager and/or quarantine officer.

• B Purpose of application and use

• 1.3. Information on the application:

  • •  Indicate whether this is a first application or a renewal of a previous application. In the case of a renewal, include a dossier reference number and expiry date and highlight any changes introduced since the first application.

  • •  Is the organism on the EPPO ‘Positive List of IBCAs’⁵?

  • •  Has an application for this organism been submitted elsewhere in Europe, or has the organism or a product containing the organism been registered elsewhere in Europe? Specify in what country and contact details, when the application was submitted and the outcome.

  • •  Is there a relation with other applications currently submitted or previously licensed with other IBCAs or beneficial organism(s) in the same product?

  • •  For what period is the permit requested (within the range allowed by the relevant NCA)?

• 1.4. Purpose of Use:

  Indicate the purpose of the application and use of the organism:

  • •  Indicate whether the application is made for (i) import for research and/or (mass) rearing or (ii) direct release.⁶ Indicate whether a release is intended in the country of application or not;

  • • When releases are intended, indicate whether the applications are for trial purposes or for full field releases, in commercial and/or classical programmes;

  • •  Type of biological control programme⁷: classical biological control (CBC), augmentative (inundative) biological control (IBC), weed biocontrol;

  • •  For direct release in field trials or for commercial release, indicate whether permanent establishment is intended (classical release) or not (augmentative release);

  • •  Provide details of area of application (e.g. protected, semi-protected glasshouse, open field, natural environment).

• 1.5. Facilities and Procedures

  The research/production facilities and procedures: describe how the risks, and the extent or probability of escape into the wild will be managed (for import of non-indigenous organisms only). This can usually be done by means of one or more waivers.

  • •  Address (physical), postal code, location (city);

  • •  For imported material, provide details of labelling, packaging and storage during transit;

  • •  Facility: describe the types of facilities used (greenhouses, laboratories, climate rooms or cabinets);

  • •  Levels of containment: do you have a permit to work with quarantine organisms under the provisions of Directive EC/95/44⁸? If not, justify why the levels of containment proposed for transport, rearing or research are appropriate to avoid escape and spread; where feasible, a contingency plan to prevent undesired environmental effects should be provided.

  • •  Quality control management system: give a description of the measures, methods and intervals to ensure quality and purity of the IBCA (species/strain), and methods for periodic control of purity and identity of mass-rearing, including Standard Operating Procedures for:

    • ∘  Life stage and numbers (amount) to be imported;

    • ∘  Methods and materials to be used for shipping (e.g. sealed container, host mummies, prey to be included, plant material included, etc.);

    • ∘  Procedures to eliminate any contaminants of the imported agent that are of concern;

    • ∘  Procedures to dispose of used research materials, including shipping materials;

    • ∘  A plan for detecting escape and undesired environmental effects;

    • ∘  Any other procedures specific to this importation (i.e. not part of standard procedures).

  • • Accreditation: is your organization certified and/or accredited for processes and/or activities (ISOs) as developed by the International Organization for Standardization.⁹ Relevant standards include ISO 9001 for ‘Quality management’ (general procedures) and ISO/IEC 17025 for ‘General requirements for competence of test and calibration laboratories’. Provide details of the ISO standard(s) and activities for which you have certification and/or accreditation.

• 1.6. Information on the target organism(s) and area of application

  • •  Name(s) of pest(s) to be controlled (order, family, genus, species and author), including weeds;

  • •  Origin of the pest(s)/weeds and the natural occurrence in the area of release;

  • •  Biology of pests: life cycle(s) of pests/weeds released against;

  • •  Crops: damage inflicted on target crops or vegetation; crops or vegetation on which releases will be made.

4 Part 2. Information for indigenous and non-indigenous IBCAs

Table 16

• 2.1. Identity and ID Confirmation

  For what species/organism is the application made? Indicate which species is involved (a single species per application) and full scientific name and taxonomy. Give an accurate identification of the IBCA or, where necessary, sufficient characterization to allow its unambiguous recognition, such as

  • Order, family, genus, species and author, and, where appropriate, sub-species, strain, or biotype; include common names and synonyms;

  • Include the name of micro-organisms directly associated with the IBCA, e.g. identity of the symbiotic bacteria in entomopathogenic nematodes.

    ID confirmation: Indicate means, methods of ID confirmation and vouchers:

  • Authority: by which expert or institute has the organism been identified?

  • By what method (morphological, molecular): if available, include a letter from a scientific expert, recognized by the NCA, stating the identity of the organism;

  • Supply evidence of deposition of voucher specimens, with identity confirmed, in a recognized collection facility (these depositions must be made before the agent is released); include the name and location of institution(s) where voucher specimens are deposited;

  • Where cultures are refreshed, confirmation of identity should be sought at regular intervals and additional vouchers should be deposited accordingly;

  • Include the accurate identity of the symbiotic bacteria associated with entomopathogenic nematodes used as an IBCA.

    • 2.2. Characterization of IBCA

      Specify life-stages, strains or taxonomic constraints:

  • General diagnostic descriptions of all life stages of the IBCA that are relevant for its use in biological control, highlighting details of any taxonomic characteristics and difficulties with the group (e.g. species complexes, cryptic species, poorly studied group);

  • Describe specific characteristics of the species/strain(s) (where relevant), such as:

    • ∘  cold-hardiness (winter survival, diapausing abilities);

    • ∘  known pesticide resistance (if yes: what resistance);

    • ∘  information on differences from the parent wild strain.

  • Where appropriate, molecular information (e.g. unique micro-satellite markers) used for diagnosis, especially for population identification, species complexes or cryptic species.

• 2.3. Origin and Distribution

  What is the immeditate source of the organism. Include details of the origin and distribution of the IBCA (species or lower taxon) as follows:

  • a) Indicate whether indigenous or non-indigenous

  • b) If field collected, provide information on collection sites and dates, including:

    • •  geographic area (approximate latitude, longitude and altitude of site);

    • •  description of the original habitat(s) and host(s) from which the collection was made.

• c) If from laboratory culture or production facility, provide information as indicated in (a) and in addition, the history of the culture stock, including:

  • the immediate source of the organism (i.e. where it is produced), giving the name and address of the manufacturer, including the location of the production facility;

  • any other source from which the culture has been collected or supplied;

  • frequency and origin of additional wild stock used to refresh laboratory cultures.

    • d) Current distribution, including:

  • Known areas of original natural distribution of the IBCA;

  • Known areas where the IBCA has been intentionally or accidentally introduced

    B Product information

    • 2.4. Product Information

      For augmentative (inundative) commercial release or classical biocontrol, briefly describe the intended use and potential benefits that may be derived.

  • Function of the IBCA (e.g. predator, parasitoid);

  • Life stage(s) of the agent(s) to be released (e.g. pupae, adults);

    For augmentative (inundative) commercial releases, the following information should be supplied:

  • Trade name of the product;

  • Method of supply and formulation (e.g. single species, interim prey, mixed species);

  • Label and container information;

  • Storage conditions (temperature, humidity, expiry date);

  • Recommended method of use (e.g. frequency and dosage of release).

    • 2.5. Product Composition

      Provide evidence that for inundative releases, the product is free from unwanted contaminants i.e. entomopathogens and hyperparasitoids, including:

  • Co-formulants: give a description of co-formulants/organic contaminants included with the IBCA (e.g. plant material, live prey or other food materials, carrier material);

  • Contaminants: give an assessment of the extent to which these should be of concern; frequency and percentage of hosts used in culture that might be present in the marketed product;

  • Any combined or contaminant organism should be separately authorised before import and/or release.

In the case of a renewal of a previously successful application (Section 1.3), or if the species or population is indigenous to the country or ecoregion, and/or imported for research or rearing only, and/or mentioned on the list of species considered safe for use in the intended area of release, no further information is required and only the submission details in Part 4A and B and Appendices (Part 5) need to be completed. For other applications, such as the release of a non-native species, the information requirements in Part 3 must be supplied.

5 Part 3. Information requirements for intentional release of a non-indigenous IBCA

A Biology and ecology

• 3.1. Information on the biology and ecology (in current area of distribution)

  Information provided below will be the main basis for the environmental risk assessment. Give a description of the biology and ecology of the IBCA, including:

  • Life cycle and number of generations per year;

  • information on developmental and reproductive biology (e.g. sexual/asexual reproduction, feeding and parasitisation habits, developmental period, reproductive potential, longevity);

  • known mechanisms of survival of extreme conditions (e.g. diapause, quiescence, migration);

  • known mechanisms of dispersal (e.g. flight capability, migratory behaviour);

  • describe the climatic conditions of areas where the IBCA is known to be native and/or where it has established following intentional or accidental introductions;

  • give information on the habitat range, including the habitat(s) where the IBCA is known to be native and/or where the IBCA is known to have established following intentional or accidental introductions (e.g. pasture, forest, scrub, etc) and known factors determining habitat selection (e.g. oviposition behaviour);

  • Give details of natural enemies, including pathogens known to attack the IBCA.

    B Assessment of risks and benefits

    Information presented in this section forms the basis for the ERA. The ERA should address the whole country within which releases will be made, with reference to regional variation that may affect risk where appropriate. Information required in this section is considered essential to an ERA, and can be acquired from published literature, company reports and/or experimentation. Include details of previous risk assessments for the same species (strain/biotype) with outcomes and other relevant information, including the country of application. The submission of available and/or generated data and subsequent assessment of environmental risks follows a tiered approach: information should be acquired and risks assessed according to the hierarchical system proposed by Van Lenteren et al. (2003, 2006), and further updated in REBECA Work Package 5. When establishment of the IBCA is very unlikely and the organisms released are predicted to die out, the subsequent fields need not be filled in, and no further risk assessments are necessary; when establishment of the IBCA is likely or necessary (e.g. in classical control), host range information is a crucial requirement for risk assessment; dispersal test results are needed when IBCAs are released in open fields and establishment is very unlikely; a summary of known direct and indirect non-target effects should always be given.

• 3.2. Safety and Health Effects

  Summarize available information on hazards to human, animal and plant health (for example, allergy, skin irritation, disease vectoring etc) by the IBCA, product or any co-formulants and measures taken to limit operator exposure, where necessary.

• 3.3. Information on Environmental Risk Assessment (ERA)

  All fields should normally be completed (but see exemptions listed below), but may be weighted differently in the evaluation of risks. Summarize the history of previous releases or introductions and the outcome of previous risk assessments, with known consequences, including non-target effects.

  • 3.3.1. Potential for Establishment

    Indicate any evidence of establishment as a result of previous releases or accidental introductions outside Europe or other IOBC/WPRS countries. Describe conditions (including extremes) affecting the IBCA’s survival and reproduction in its current distribution.

    Information on physical constraints, such as:

    • •  Climatic similarities/differences between area of current distribution and area of intended release (e.g. temperature, altitude, humidity, day length, etc.);

    • •  Probability of temporary survival;

    • • Ability to survive and reproduce at temperatures and humidities outside the normal range (e.g. cold tolerance, overwintering ability); lower and upper temperature thresholds for development and survival; ability to enter diapause and/or overwinter (include test results);

    • •  Other physiological and behavioural mechanisms for surviving extreme conditions;

    • •  Dispersal potential (where known);

      Information on resource constraints, such as:

    • • Availability and utilization of suitable hosts (target and non-target organisms) for short-term or long-term survival;

    • •  Availability of suitable habitat, vegetation and plant food resources.

      Indicate any evidence of establishment as a result of previous releases and/or accidental introductions outside Europe.

      When outdoor establishment of the IBCA is very unlikely and the organisms released are predicted to die out rapidly, the subsequent fields need not be completed, and no further risk assessments will be necessary; when outdoor establishment of the IBCA is likely or necessary, host range information must be supplied.

  • 3.3.2. Host Range Assessment

    When establishment is likely and/or required, provide available information on recorded effects on non-target organisms, including:

    • •  A list of known hosts other than the target pest(s) and potential of the IBCA to utilize non-target host organisms living on wild or cultivated plants;

    • •  A list of non-target organisms that have previously been tested, including unrelated non-target hosts, including pollinators, and threatened and endangered species; indicate hosts that were not accepted in such tests;

    • •  Procedures used to determine host range (e.g. phylogenetic relatedness, experimentation) and methods used for host-range testing (e.g. experimental design, test conditions, rearing methods for non-target species, life-stages tested etc);

    • •  Possible direct effects on plants: describe possible direct effects of the IBCA on the host plant(s) of the target pest and on plant hosts of non-target species.

• 3.3.3. Dispersal

  • Indicate potential direct (inundative) effects of mass-releases into open fields to neighbouring non-target hosts and habitats;

    Direct effects of dispersal are considered for both indigenous and non-indigenous IBCAs where relevant to the direct environment of release. Dispersal test results are not required for glasshouse releases, but should be provided when IBCAs are released in open fields or structures that do not prevent escape (e.g. polytunnels) and long term establishment is very unlikely.

    • 3.3.4. Additional Information on Direct and Indirect Non-target Effects

      Describe the history of previous releases or accidental introductions, with known consequences, including non-target effects. Indicate any other possible specific non-target effects, such as:

  • Competition with, or displacement of, indigenous natural enemies in the area of intended release;

  • Other constraints on the presence of natural enemies, including transfer of pathogens, of the released IBCA;

  • Presence of natural enemies, including pathogens, that may affect establishment of the IBCA

    A summary of known direct and indirect non-target effects should always be given, irrespective of whether host range and/or dispersal have been assessed. This section should also include conclusions on the risks associated with the intended release.

    • 3.4. Efficacy and Benefits of the IBCA and Proposed Release

      Provide relevant information on:

  • Anticipated contribution to the control of the target pest(s) and weeds;

  • Estimated economic benefits (crop specific) of the IBCA;

  • Possible environmental benefits, e.g. beneficial effects of release of the IBCA compared with current control methods;

  • Method(s) to determine efficacy and, when required by the NCA, results of efficacy trials.

6 Part 4. Submission of Forms and Signature

A Submission details

• 4.1. Appendices

  Check your application for completeness in the following areas:

  • Information requirements (dossier)

  • References, other literature and overview of information used in preparation of the dossier: include copies of relevant articles, chapters or reports in an appendix to the application documents;

  • Identification of applicant: ID-card or passport;

  • Chamber of Commerce copy;

  • Authorization for payment of fees;

  • Letter from a scientific expert, recognized by the NCA, confirming identity of the organism;

  • Evidence of deposition of voucher specimens, with identity confirmed, in a recognized collection facility (these depositions must be made before the agent is released); include the name and location of institution(s) where voucher specimens are deposited

  • In case of import for research and/or rearing, include a map of the facilities;

  • Any other information that is relevant to the application.

• 4.2. Where to Submit the Application

  Address details of the NCA

  B Agreement

  • 4.3. General Safeguards

    The applicant or authorized user undertaking the release proceeds under the conditions of the authorization for release, taking into account the following requirements:

    • •  All appropriate safety procedures should be put in place.

    • •  Any relevant information on adverse effects which might relate to the released IBCA should be reported immediately to the NCA.

    • • Information on sites and dates of supply or release of the IBCA should be made available to the NCA if requested.

    • • Information requirements have been supplied according to the most recent knowledge, and that the conditions made by the NCA will be respected.

  • 4.4. Signature Details

• • Date

• • Applicant’s name

• • Signature

All information and documents submitted for a licence application (dossier) will be regarded as ‘commercial in confidence’ by the NCA. The Environmental Risk Assessment and decision will be based on data and documents submitted for that specific licence application only.

7 Part 5. Appendices

1 Appendix 3: Summary of Methods to be Used in the Environmental Risk Assessment of Invertebrate Biological Control Agents

Introduction

This appendix should be read with reference to Fig. 16.1 in Chapter 16 and other information presented in the chapter, particularly the order of testing in an environmental risk assessment, flexible routes through the ERA system, the granting of waivers (exemptions) and taxon-specific issues. In most cases the ERA would follow the order of ‘Establishment’, ‘Host Range’ and ‘Dispersal’, although there are situations in which some of these assessments could be omitted, by-passed or conducted in a different order (for examples, see text in chapter). This system and methods have been devised primarily for arthropod biological control agents, but the principles are applicable to entomopathogenic nematodes, as summarized in the Appendix.

Establishment

• 1. Long term establishment of a non-native species has two main requirements: (i) ability to survive in the climate in the area/country of release, and (ii) access to a food resource – usually, ‘wild prey’ (where ‘prey’ is synonymous with ‘host’), which could include established ‘exotic’ species. It is recommended that both of these requirements are assessed (though not necessarily ‘tested’), as this may identify some species that are ‘climatically suited’ for establishment but unable to establish because of the absence of any acceptable wild prey.

• 2. In general, an ability to diapause increases the likelihood of winter survival, and in turn, longer term establishment. For this reason, ability to diapause should be investigated as a matter of routine in inundative biocontrol, especially where source populations are collected from different countries or different regions within countries. Information on diapause may be available in the literature or acquired by experimentation. Diapause induction stimuli vary between species but in most cases winter diapause can be induced by a 12:12 LD cycle at 15°C (and often by 12:12 LD at 20°C).

  Key point: Ability to diapause should be assessed as a matter of routine prior to other ‘tests’ for establishment for species intended for inundative release (glasshouse or field), where an inability to diapause would be a desirable feature. Diapause studies are less important for classical control where establishment is normally a requirement for success.

• 3. Climatic suitability (most often, overwintering ability) can be assessed by the system developed by Bale and co-workers (Hatherly et al., 2005) in which laboratory survival at 5°C is a reliable predictor of duration of field survival in winter, in northern European countries or regions with a winter climate similar to the UK. The system is now based on 8 (mainly predatory) species, which all ‘conform’ within a strongly correlated relationship. This approach enables species for inundative biocontrol to be categorised as ‘safe’ (die out within about 4 weeks of release), ‘marginal’ or ‘likely to establish’ (can survive for entire winter). Further analyses should be conducted to identify as far as possible ‘survival time limits’ for each category, and the extent of the ecoregion to which the data could be applied. Also, as most of the species examined so far are predators, further studies are required to assess the wider applicability of this system to parasitoids. It should also be noted that whereas as some predatory species have a cold hardy stage of the life cycle that can survive throughout winter, there are likely to be some parasitoid species that are not so long-lived, but are sufficiently cold hardy to complete one or more generations in winter and thus have the same or greater establishment potential. As those species which ‘die out quickly’ are usually unable to survive below their developmental threshold (often in the range of 8–10°C for species of tropical origin), the developmental threshold might be an additional predictor of establishment. This could be investigated as the data are usually available in the literature, but some caution is required as reported thresholds may vary depending on regional variations in different source populations, stage specific differences, and differences related to different prey-host plant combinations used in experiments and in commercial production.

  Key point: When experimental data on establishment are required, it is proposed that a laboratory assessment of survival at 5°C is appropriate to predict field survival, particularly for weakly cold tolerant species. Companies could usefully indicate to which countries or ecoregions such data would apply.

• 4. The species that pose interpretational difficulties are those in the ‘marginal’ zone that can survive for 1–2 months but not entire winters, and cannot reproduce in winter. For such species, it would be relatively easy to assess their acute lethal temperature and compare this with regularly occurring minimum temperatures in areas of intended release. However, as the effect of cold stress is determined by both the temperature and the duration of exposure, the reliability of this ‘quick test’ for ‘marginal’ species requires further evaluation.

  Any ‘climate survival’ test should include different life cycle stages (unless there is a known overwintering stage), with and without an acclimation treatment, and where appropriate, with access to a food (host or prey) resource.

  Key point: For species that are predicted (or shown) to die out after brief periods of winter low temperatures, no further risk assessments are necessary, other than a consideration of direct and indirect effects, as for a native species.

• 5. The second requirement for establishment is availability of one or more species of wild prey (which, depending on the climate, may be target or non-target species). The ability of the candidate agent should be assessed on one, or a small number, of commonly available wild prey that are phylogenetically related to the target species. With the benefit of experience it may be possible to produce a ‘recommended list’, but as an example, if the glasshouse target was a species of whitefly, then the cabbage whitefly Aleyrodes proletella would be an appropriate wild prey. For most non-native biocontrol agents there are likely to be suitable wild prey, but if a candidate species did not feed on one or more close relatives of the target, this might be an indication of host specificity, and would therefore be valuable information in the overall risk assessment. In this part of the establishment assay, the response of the control agent should be recorded in terms of attack (attempt to feed or oviposit), death of the prey, and ability of the agent to develop on the wild prey and produce reproductively viable adults.

Host Range

• 6. The second aspect of risk assessment of inundatively released agents is host range, but this would be the first area of investigation for a classical control agent (see Introduction to this appendix). There have been a number of studies and recommendations on host range testing. It is recommended that the testing scheme for arthropod biocontrol agents proposed by van Lenteren (2006b) should be adopted and non-target species for host specificity testing selected as described by Kuhlmann et al. (2006). Testing schemes for weed biocontrol have been reviewed by Sheppard et al. (2005), and the selection of non-target species follows recommendations made by Wapshere (1974).

  Key point: Species selected as ‘test’ prey and hosts are used to obtain an indication of the likely host range, not a precise list of non-target species that are accepted or rejected. For this reason, the selected list should be representative of different taxonomic groups rather than a particular country. Ideally, the same list, or at least a similar one, should applicable across Europe.

• 7. Host range testing can be an expensive exercise, beyond the financial limitations of even the largest companies. For this reason, it is proposed that host range testing should be conducted in two stages. It is unwise to be prescriptive about the exact number of species to be used in each stage, but typically this could be 3 species in stage 1 and a further 6 species in stage 2 of an arthropod biocontrol programme. This would allow companies to decide at stage 1 whether to continue with further host range testing.

  Key point: The identity and number of species to be included in host range tests should ideally be discussed with experts and agreed with the regulator prior to any experimentation.

  Stage 1 assessment should include a phylogenetically close relative of the target prey or host (such as the species used in the establishment assay above), a second close relative, and a third species that is taxonomically distinct but commonly available outdoors, including during winter when appropriate to the seasonal biology of the agent. Data recorded should be attack, death of prey or host and development to adult as with the establishment assay.

• 8. Where the Stage 1 test indicates some level of specificity (e.g. only the phylogenetically related species are accepted as prey or hosts), testing should proceed to stage 2. For arthropod biocontrol, the system proposed by Kuhlmann et al. (2006) is recommended in which non-target species are selected from three categories: 1. Phylogenetically related; 2. Occurs in the same ecological niche; 3. Unrelated ‘safeguard’ species.

• 9. A number of studies have compared the physiological (‘apparent’) host range of some parasitoids, and the ‘ecological’ host range that is observed in nature. Invariably, laboratory assessments in which hosts are offered to natural enemies in ‘no choice’ tests overestimates the natural host range. The stepwise procedure proposed by Van Lenteren et al. (2006b) is recommended as the method that should be used for arthropod biocontrol to make an estimation of the range of non-target species attacked under field conditions.

  If the host/prey is accepted in the first two steps (conducted in small arenas), the step 3 test should be carried out in contained environments such as large cages, in which prey or hosts feed on growing plants and the agent is able to move freely around the cage. It is recommended that three treatments are compared with appropriate replication: 1. Target species alone (control); 2. Non-target alone; 3. Target and non-target together.

  Key point: If acceptance of non-target hosts is observed in no-choice tests, a further test needs to include direct comparison of the acceptance and development on non-target species when the target species is simultaneously available.

Dispersal

• 10. In general, the need to assess dispersal will be restricted to a limited number of candidate agents for inundative biocontrol. If it is clear that a species can establish in the release environment, it should be assumed that dispersal will occur – the unknown factors being ‘how soon’ and ‘how far’, and these are both difficulty to quantify on a ‘pan-European’ scale. However, there are circumstances in which dispersal may be limited (flightless species), and such information should be provided in a dossier.

  Key point: Dispersal should not be tested in species that can establish in the release environment.

• 11. If no establishment is predicted, any effects on the wider environment will be transient and generally restricted to the ‘summer season’. It is recommended that dispersal should not be assessed in species that are used exclusively in glasshouses where any escapes will involve low numbers of individuals that will have minimal impact on the neighbouring species and ecosystem before they die out.

  Key point: Dispersal should be tested only when agents are released into open fields or structures that do not restrict escape.

• 12. The impact of an ‘open field’ release where there is no prospect of survival through winter will depend on the numbers released and dispersal distances, and the proximity of the release area to sites of special scientific interest, such as nature reserves. Dispersal data are generally difficult to obtain but a description of methods by which to assess dispersal for inundatively released biological control agents is provided by Mills et al. (2006). It is also recommended that a database of information should be created from the literature and experimental studies to provide ‘typical dispersal distances’ for different taxonomic groups commonly used in biocontrol. Companies should have the discretion to provide information on atypical species with limited dispersal ability.

Direct and Indirect Effects

• 13. Direct and indirect effects are a summary of information gained from the available literature. When such information is not readily available, these effects may be estimated by ‘expert knowledge’ or generated from the data on establishment, host range and dispersal in the ERA. Examples of direct effects would include effects on non-target species and on other trophic levels (such as intraguild predation and plant feeding damage), hybridization and enrichment and vectoring (van Lenteren et al. 2003; Bigler et al. 2006). Indirect effects are those that occur when there is no direct interaction between the control agent and non-target species, such as competition and competitive displacement (see van Lenteren et al. 2003; Bigler et al. 2006). Indirect effects are difficult to quantify, but are likely to be related to the scale of the direct effects.

• 14. In situations where winter survival of the candidate agent for inundative biocontrol has been demonstrated in the establishment experiments (or seems likely to occur) and where the species is known or shown to be polyphagous, a company may decide that further investment in host range or other forms of testing would not be cost effective, as the dossier may not lead to a successful licence application. In such situations, a company could prepare a dossier describing a ‘worst case scenario’ that might arise from a release and provide relevant information for a ‘risk-benefit’ analysis compared with other available methods of control. In effect, although the biological control agent may pose some risk, this may be less than for other control options. A problem with this approach is that there may be difficulties in obtaining reliable comparative data for the alternative method(s) of control. However, there are examples of previously released species that have survived in the northern European climate, and are known to be polyphagous, but as yet, have not had any detectable impact on native species or ecosystems. A risk assessment for such species evaluated under current regulatory guidelines would almost certainly lead to a ‘licence rejection’ when considered in isolation, but the species might be the best option in comparative terms.

  Key point: For polyphagous agents with establishment potential, companies should have the option to submit a dossier containing information on the risks and benefits of the proposed release compared with other possible controls. This information would be evaluated by the regulator as part of the ERA.

• 15. Direct and indirect effects of classical biological agents should be addressed in pre-release studies, because establishment of such species is essentially irreversible. Additionally, negative direct effects of classical biocontrol agents on non-target prey or hosts have become a major issue in this method of control.

Nematodes

• 16. The proposed ERA should include entomopathogenic nematodes (EPNs), allowing for the development of appropriate methods and modification to the order of testing as appropriate. On the basis of available information EPNs (i) have very limited potential to cause non-target effects, and (ii) should be included within the same ERA framework that is applied to insects and mites, but with the recommendation that data on establishment, dispersal, host range and indirect and direct effects would not normally be necessary because of the limited potential of EPNs to disperse or persist at the site of application. The remote risk related to the use of Heterorhabditis indica can be excluded by a precise identification of its associated symbiotic bacterium.

Related Issues:

• 17. It is likely that companies will want to carry out efficacy trials and ERA experiments simultaneously to minimize the time between product development and commercial release. Some of the efficacy work needs to be conducted under commercial or semi-commercial conditions (to determine effectiveness of agent on different crops, release rates etc), but this would pose some risk in species with the potential to establish. It was recommended that ‘establishment potential’ should be assessed before any commercial scale efficacy trials. In situations where there is no prospect of establishment in the local environment, companies should be able to conduct efficacy trials under outdoor or open field conditions. Where some establishment is possible or likely, the location and biosecurity of efficacy trials should be discussed with the regulator. As a general principle, companies should conduct such trials in a contained facility (large cage, glasshouse), taking all reasonable effort to prevent escape, in sites that are geographically isolated from areas of ‘scientific sensitivity’, and with regular monitoring in the immediate vicinity of the trial to detect any occurrence of the agent outside of the enclosed environment. When such escapes are observed, the trial should be terminated immediately and all plants and invertebrate material destroyed. Similarly, at the end of the trial, all plants and pests/control agents should be destroyed. These conditions should be applied to all researchers involved in biocontrol research, including universities and research institutes.

  Key point: Establishment potential of inundative biocontrol agents should be assessed prior to commercial scale efficacy trials. For species with no ability to establish in the climatic area of the trial, experiments can be conducted under ‘open field’ conditions if appropriate. When establishment is possible or likely, an appropriate level of biosecurity should be adopted in discussion with the regulatory authority.